Human CKMT1A Knockout HEK293T Cell Line

Storage Condition	Upon arrival, the vial should be stored in liquid nitrogen vapor phase. Storage buffer: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether
Shipping Condition	Shipped on dry ice.

Parental Cell Line	HEK293T
Passage Number	<20
Viability	~80%
Mutation Description	CRISPR/Cas9 system is used to generate knockout cell lines. Homozygous: 1 bp insertion in exon 4.
Adherent / Suspension	Adherent
Cell Type	Epithelial Cells
Mycoplasma Free	Yes
Knockout Validation	Sanger Sequencing
Cell Medium	Culture medium: DMEM (High Glucose) + 10% FBS. Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Initial Handling Guidelines	1. Thaw the vial in 37ºC water bath approximately 1-2 minutes. 2. Transfer the cell suspension to a 15 ml/50 ml conical sterile polypropylene centrifuge tube containing pre-warmed culture medium. Wash vial with an additional culture medium to collect remaining cells, and centrifuge at 201 × g (rcf) for 5 minutes at room temperature. 3. Resuspend the cell pellet in 5 ml pre-warmed culture medium and count cell number. Seed cells in an appropriate cell culture flask at a density of 2×10⁴ cells/cm². This should allow for confluency within 48 hours. 4. Incubate the culture at 37ºC incubator with 5% CO₂. Cultures should be monitored daily.
Subculture Guidelines	Cells should be passaged when they have achieved 80-90% confluence. A partial media change 24 hours prior to subculture may be helpful to encourage growth.
Description	Human wild-type HEK293T cell line as the recommended control.

Target Function	Creatine kinase MT is responsible for the transfer of high energy phosphate from mitochondria to the cytosolic carrier, creatine. Overexpression of ubiquitous mitochondrial creatine kinase is involved in many malignant cancers with poor prognosis.
Cellular Localization	Cell membrane and mitochondrial.
Target Sequence Similarities	Belongs to the creatine kinase isoenzyme family.